Open-source cell culture automation system with integrated cell counting for passaging microplate cultures

Courville, G.; Vaid, S.; Toruno, A.; Lebel, P.; Cabrera, J.; Raghavan, P.; Jacobsen, A.; Bell, G. R.; Leonetti, M. D.; Gomez-Sjöberg, R.

doi:10.1101/2024.12.27.629034

bioRxiv · 10.1101/2024.12.27.629034

Open-source cell culture automation system with integrated cell counting for passaging microplate cultures

Courville, G.·Vaid, S.·Toruno, A.·Lebel, P.·Cabrera, J.·Raghavan, P.·Jacobsen, A.·Bell, G. R.·Leonetti, M. D.·Gomez-Sjöberg, R.

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Abstract

Tissue culture in 96-well microplates is conventionally a tedious, highly manual process sensitive to individual technique and experimenter error. Here, we describe the Automated Cell Culture Splitter (ACCS), a system for passaging plates of adherent or suspension cells, for routine culture maintenance or specialized applications such as seeding plates for microscopy. The system is built around the Opentrons OT-2 liquid handling robot and incorporates a novel on-deck imaging-based cell counter which allows it to compensate for density disparities across a source plate and control the number of cells seeded on a per-well basis. We find this solution can cut hands-on time by 61% and the results compare favorably to our existing manual cell culture processes in terms of both seeding density precision and bio-logical outcomes, achieving a control of seeding density with a well-to-well coefficient of variation (CV) under 11%. The system is designed to be adaptable and an accessible entry point into automation for high-throughput cell culture; to that end, all of the source code and hardware designs are released under open source licenses.

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Courville, G., Vaid, S., Toruno, A., Lebel, P., Cabrera, J., Raghavan, P., Jacobsen, A., Bell, G. R., Leonetti, M. D., Gomez-Sjöberg, R.. 2024-12-27. Open-source cell culture automation system with integrated cell counting for passaging microplate cultures. https://doi.org/10.1101/2024.12.27.629034

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